Regulatory role of charged clusters in the N-terminal domain of BetP from Corynebacterium glutamicum
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Izabela Waclawska
Abstract
The trimeric transporter BetP counteracts hyperosmotic stress by a fast increase in transport rate in order to accumulate the compatible solute betaine. The positively charged α-helical C-terminal domain acts as an osmosensor perceiving the increase in the internal potassium (K+) concentration. A second, still unidentified stimulus originates from stress-induced changes in the physical state of the membrane and depends on the amount of negatively charged lipids. BetP possesses a 60-amino acid (aa)-long negatively charged N-terminal domain, which is predicted to adopt a partly helical fold affecting osmoregulation by an unknown mechanism. It is assumed that the C-terminal domain, the N-terminal domain, and negatively charged lipids interact during stress sensing and regulation. Here, we have investigated the regulatory role of negatively charged clusters in the N-terminal domain. We identified one cluster, Glu24Glu25, to be crucial for osmoregulation. Cross-linking studies revealed an interaction between the C- and N-terminal domains of adjacent protomers modulating transport activation. A regulatory partner-switching mechanism emerges in which the C-terminal domain changes its interaction with the N-terminal domain of its own promoter and negatively charged lipids to an interaction with the N-terminal domain of an adjacent protomer and lipids bound to the central cavity of the BetP trimer.
Acknowledgments
We would like to thank Reinhard Krämer for important and helpful discussions and Caroline Koshy for help with the mutant design. This work was supported by the SFB 807: ‘Transport and Communication across Biological Membranes’.
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©2015 by De Gruyter
Artikel in diesem Heft
- Frontmatter
- Meeting Report
- Membrane Transport and Communication in Frankfurt: Speakers’ Summary – Highlights
- HIGHLIGHT: MEMBRANE TRANSPORT AND COMMUNICATION
- Structure, function, evolution, and application of bacterial Pnu-type vitamin transporters
- Team work at its best – TAPL and its two domains
- The volume-regulated anion channel is formed by LRRC8 heteromers – molecular identification and roles in membrane transport and physiology
- Extending native mass spectrometry approaches to integral membrane proteins
- Functional diversity of the superfamily of K+ transporters to meet various requirements
- The structure of Na+-translocating of NADH:ubiquinone oxidoreductase of Vibrio cholerae: implications on coupling between electron transfer and Na+ transport
- Hybrid rotors in F1Fo ATP synthases: subunit composition, distribution, and physiological significance
- Homeostatic control of biological membranes by dedicated lipid and membrane packing sensors
- The transporter associated with antigen processing: a key player in adaptive immunity
- The pseudo-atomic structure of an RND-type tripartite multidrug efflux pump
- The assembly and disassembly of the AcrAB-TolC three-component multidrug efflux pump
- A universal mechanism for transport and regulation of CPA sodium proton exchangers
- Biosynthesis of membrane dependent proteins in insect cell lysates: identification of limiting parameters for folding and processing
- Fluorescence and excited state dynamics of the deprotonated Schiff base retinal in proteorhodopsin
- Regulatory role of charged clusters in the N-terminal domain of BetP from Corynebacterium glutamicum
- The contribution of methionine to the stability of the Escherichia coli MetNIQ ABC transporter-substrate binding protein complex
- The ABC exporter MsbA probed by solid state NMR – challenges and opportunities
- Functional properties of LptA and LptD in Anabaena sp. PCC 7120
Artikel in diesem Heft
- Frontmatter
- Meeting Report
- Membrane Transport and Communication in Frankfurt: Speakers’ Summary – Highlights
- HIGHLIGHT: MEMBRANE TRANSPORT AND COMMUNICATION
- Structure, function, evolution, and application of bacterial Pnu-type vitamin transporters
- Team work at its best – TAPL and its two domains
- The volume-regulated anion channel is formed by LRRC8 heteromers – molecular identification and roles in membrane transport and physiology
- Extending native mass spectrometry approaches to integral membrane proteins
- Functional diversity of the superfamily of K+ transporters to meet various requirements
- The structure of Na+-translocating of NADH:ubiquinone oxidoreductase of Vibrio cholerae: implications on coupling between electron transfer and Na+ transport
- Hybrid rotors in F1Fo ATP synthases: subunit composition, distribution, and physiological significance
- Homeostatic control of biological membranes by dedicated lipid and membrane packing sensors
- The transporter associated with antigen processing: a key player in adaptive immunity
- The pseudo-atomic structure of an RND-type tripartite multidrug efflux pump
- The assembly and disassembly of the AcrAB-TolC three-component multidrug efflux pump
- A universal mechanism for transport and regulation of CPA sodium proton exchangers
- Biosynthesis of membrane dependent proteins in insect cell lysates: identification of limiting parameters for folding and processing
- Fluorescence and excited state dynamics of the deprotonated Schiff base retinal in proteorhodopsin
- Regulatory role of charged clusters in the N-terminal domain of BetP from Corynebacterium glutamicum
- The contribution of methionine to the stability of the Escherichia coli MetNIQ ABC transporter-substrate binding protein complex
- The ABC exporter MsbA probed by solid state NMR – challenges and opportunities
- Functional properties of LptA and LptD in Anabaena sp. PCC 7120
